CN103655744A - Preparation method for Fukean tablet and application of Fukean tablet in preparation of drug used for inhibiting proliferation of mouse lymphoma cell YAC-1 - Google Patents
Preparation method for Fukean tablet and application of Fukean tablet in preparation of drug used for inhibiting proliferation of mouse lymphoma cell YAC-1 Download PDFInfo
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- IVTMALDHFAHOGL-UHFFFAOYSA-N eriodictyol 7-O-rutinoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C=C3C(C(C(O)=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 IVTMALDHFAHOGL-UHFFFAOYSA-N 0.000 description 9
- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 description 9
- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 description 9
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 description 9
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- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
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- WEEMDRWIKYCTQM-UHFFFAOYSA-N 2,6-dimethoxybenzenecarbothioamide Chemical compound COC1=CC=CC(OC)=C1C(N)=S WEEMDRWIKYCTQM-UHFFFAOYSA-N 0.000 description 1
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- Medicinal Preparation (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the technical field of traditional Chinese medicine and particularly relates to a preparation method and application of a Fukean tablet. The Fukean tablet is prepared from 1000 g of Jasminum elongatum, 1000 g of Chinese knotweed herb, 670 g of Ilex rotunda Thunb, 330 g of plantago and 330 g of pomegranate rind through microwave extraction and macroporous resin adsorption; so total flavonoid content is greatly improved. The invention also provides application of the Fukean tablet in preparation of a drug used for inhibiting proliferation of the mouse lymphoma cell YAC-1.
Description
Technical field
The invention belongs to technical field of Chinese medicines, be specifically related to a kind of preparation method of fukean tablet and suppress the application in mouse lymphoma cell YAC-1 cell proliferation medicine in preparation.
Background technology
Fukean tablet standard No. WS-10256(ZD-0256)-2002, be recorded in national standard for traditional Chinese medicines compilation internal medicine taste fascicle.By Herba Oxalidis strictae 1000g, Herb Polygoni Chinensis 1000g, Cortex Ilicis Rotundae 670g, Herba Plantaginis 330g, Pericarpium Granati 330g, as crude drug, made, there is clearing away heat-damp and promoting diuresis, convergence, the effect of pain relieving.The stomachache, diarrhoea, the vomiting that for acute gastroenteritis, dyspepsia, cause.
In prior art, not yet there is fukean tablet aspect extraction preparation, adopting the report of microwave technology and macroporous resin adsorption extractive technique, and decocting in water alcohol extraction, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, be inconvenient to take, had a strong impact on this product and applied clinically.
Summary of the invention
In order to solve above-mentioned technical problem, the invention provides a kind of preparation method of fukean tablet.
Another object of the present invention is to provide a kind of fukean tablet to suppress the application in mouse lymphoma cell YAC-1 cell proliferation medicine in preparation.
The object of the invention is to realize by following scheme:
A kind of preparation method of fukean tablet, by Herba Oxalidis strictae 1000g, Herb Polygoni Chinensis 1000g, Cortex Ilicis Rotundae 670g, Herba Plantaginis 330g, Pericarpium Granati 330g makes as crude drug, it is characterized in that, described preparation method is comprised of the following step: get upper gomi herbs, pulverize, 65% ethanol that adds 10L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600-800W, extract 2 times, each 5-10 minute, combining extraction liquid, concentrated, be added on Diaion HP-10 macroporous adsorptive resins, 65% ethanol elution, collect 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, microwave extraction thing is mixed and adds starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.3g.
In the preparation method of above-mentioned fukean tablet, described microwave extracting power is 700W.
In the preparation method of above-mentioned fukean tablet, the each extraction time of described microwave extracting is 8 minutes.
Above-mentioned fukean tablet suppresses the application in mouse lymphoma cell YAC-1 cell proliferation medicine in preparation.
In prior art, fukean tablet is oral, one time 4,3 times on the one.Fukean tablet dosage is large.The every heavy 0.3g of fukean tablet that adopts the inventive method to be prepared into only needs 2 at every turn, within 1st, takes 3 times.Greatly reduced dose having under the condition of more active component.This conclusion can be by following evidence.
The comparison of general flavone content in fukean tablet prepared by test one, distinct methods
L, instrument and reagent fukean tablet of the present invention: press embodiment 1 method preparation, use 3330g crude drug, make 500 through extracting, every heavy 0.3g.Former fukean tablet, according to WS-10256(ZD-0256)-2002 standard method preparations, in contrast.Visible-ultraviolet light microwell plate detector (U.S. MD company model: SPECTRAMAX190); METTLER AE240 electronic analytical balance; Control substance of Rutin (Nat'l Pharmaceutical & Biological Products Control Institute).
2, method
The preparation precision of reference substance solution takes at 120 ℃ of drying under reduced pressure appropriate to the control substance of Rutin of constant weight, adds 60% ethanol and makes every 1ml containing the solution of 0.1mg, obtains.
The preparation precision of standard curve measures reference substance solution 1.0ml, 2.0ml, 3.0ml, 4.0ml, 5.0ml, puts respectively in 25ml measuring bottle, respectively add 10% sodium nitrite solution 0.3ml, mix, place 6 minutes, add 10% aluminum nitrate solution 0.3ml, mix, place 6 minutes, add 4% sodium hydroxide solution 4ml, shake up, add 30% ethanol to scale, shake up, place 15 minutes; Take corresponding solution as blank.According to spectrophotography (appendix V B of Chinese Pharmacopoeia version in 2000) test, at 510nm wavelength place, measure trap, take trap as vertical coordinate, concentration is abscissa, drawing standard curve.
Algoscopy is got this product under weight differential item, remove coating, porphyrize, (fukean tablet prepared of the inventive method is got 0.125g in sampling, former fukean tablet takes 0.2g), accurately weighed, put in round-bottomed flask, add methanol 25ml, shake up, reflux 1 hour, take out, be cooled to room temperature, filter, filter paper adds methanol 25ml again together with residue, reflux 1 hour, take out, cooling, filter, merging filtrate, methanol 15ml gradation washing for residue, washing liquid and filtrate merge, evaporate to dryness, residue adds 60% alcoholic solution to be made to dissolve, be transferred in 50ml measuring bottle and be diluted to scale, shake up.Precision measures 1ml, and method under sighting target directrix curve preparation, from " putting respectively in 25ml measuring bottle ", is measured trap in accordance with the law, from standard curve, reads the weight of rutin in need testing solution, calculates, and obtains.
3, result
Result shows, in fukean tablet of the present invention, the content of total flavones is counted 80-100mg/ sheet with rutin; And the content of total flavones is counted 10.54mg/ sheet with rutin in former fukean tablet, doubly, in the situation that dose reduces, general flavone content improves a lot the 8-10 that every rutin content is equivalent to former tablet content.
Above-mentioned research shows, the fukean tablet that adopts preparation method of the present invention to prepare, active constituent content is far away higher than WS-10256(ZD-0256) fukean tablet prepared of the method recorded of-2002 standards.
The specific embodiment
Below in conjunction with specific embodiment, the present invention is further described, so that those skilled in the art more understands the present invention, but this should be interpreted as to the scope of the above-mentioned theme of the present invention only limits to following example, all technology realizing based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Herba Oxalidis strictae 1000g, Herb Polygoni Chinensis 1000g, Cortex Ilicis Rotundae 670g, Herba Plantaginis 330g, Pericarpium Granati 330g, 65% ethanol that adds 10L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 700W, extract 2 times, each 8 minutes, combining extraction liquid, concentrated, be added on Diaion HP-10 macroporous adsorptive resins, 65% ethanol elution, collect 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, microwave extraction thing is mixed and adds starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.3g.
After testing, the content of finished product total flavones is counted 98.92mg/ sheet with rutin
Embodiment 2
Get Herba Oxalidis strictae 1000g, Herb Polygoni Chinensis 1000g, Cortex Ilicis Rotundae 670g, Herba Plantaginis 330g, Pericarpium Granati 330g, by gomi herbs, pulverize, 65% ethanol that adds 10L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600W, extract 2 times, each 5 minutes, combining extraction liquid, concentrated, be added on Diaion HP-10 macroporous adsorptive resins, 65% ethanol elution, collect 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, microwave extraction thing is mixed and adds starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.3g.
After testing, in finished product, the content of total flavones is counted 82.16mg/ sheet with rutin.
Embodiment 3
Get Herba Oxalidis strictae 1000g, Herb Polygoni Chinensis 1000g, Cortex Ilicis Rotundae 670g, Herba Plantaginis 330g, Pericarpium Granati 330g, by gomi herbs, pulverize, 65% ethanol that adds 10L, drop in microwave extracting apparatus and carry out microwave extracting, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 800W, extract 2 times, each 10 minutes, combining extraction liquid, concentrated, be added on DiaionHP-10 macroporous adsorptive resins, 65% ethanol elution, collect 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, microwave extraction thing is mixed and adds starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.3g.
After testing, in finished product, in finished product, the content of total flavones is counted 90.44mg/ sheet with rutin.
Embodiment 4: fukean tablet suppresses the experimentation data of mouse lymphoma cell YAC-1 cell proliferation
1. experiment material
1.1 experiment cell strains
Mouse lymphoma cell YAC-1 cell, Shandong University's laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: fukean tablet of the present invention: press embodiment 1 method preparation.
Medicinal liquid liquid storage: take 100mg fukean tablet, be dissolved in 5ml dehydrated alcohol, 0.2 μ m filter filters, 500 μ ldoff pipe subpackages ,-20 ℃ of storages, 0.2 μ m filter filters dehydrated alcohol in order to the use of matched group simultaneously.
1.3 experiment reagent
DMEM (Cat.No.12100-061Lot.No.758137 of GIBCO company); Hyclone (Lot.No.100419 of Tian Hang bio tech ltd, Zhejiang); NaHC0
3(Cat.No.11810-033Lot.No.1088387 of Shanghai Jiu Yi chemical reagent company limited); Trypsin(AMRESCO company); EDTA(AMRESCO company); Penicillin G Sodium Salt(AMRESCO company 1); Streptomycin Sulfate (AMRESCO); Dehydrated alcohol (Zibo Ya Dulan Trade Co., Ltd.); MTT (Biosharp lot number: 0793): the autogamy of PBS(laboratory);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DMIL); Visible-ultraviolet light microwell plate detector (U.S. MD company model: SPECTRAMAX190); C0
2incubator (FORMA model: 3111); (safe and sound company of Su Jing group manufactures model to super-clean bench: SW-CJ-ZFD); Pure water instrument (U.S. Sprlng company model: S/N020579); Accurate pipettor (French Gilson Inc model: P2); Electronic balance (German Sai Duolisi company limited model: BT323S); Full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020); Table electrothermal air dry oven (Shanghai accurate experimental facilities company model: DHG9123A); Refrigerator (Siemens Company's model: KG18V21TI); Liquid nitrogen container (CBS model: 2001); Low speed centrifuge (Anting Scientific Instrument Factory, Shanghai's model: KA-1000); 0.2 μ m filter (MILLIPORE model: SLGP033RB); 1cm culture dish (NEST company), 96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipet, Tips are some.
2. experimental technique
1) YAC-1 cell uses DMEM+10%FBS in 37 ℃, 5%C0
2carry out cellar culture (10cm culture dish), when Growth of Cells is during to logarithmic (log) phase, collecting cell, discards culture fluid, PBS fine laundering 3 times, add 3ml0.25% trypsin-0.04%EDTA, after 37 ℃ of digestion 2min, add wherein 5ml complete medium neutralization reaction, after piping and druming cell, proceeded in centrifuge tube, the centrifugal 5min of 1000rpm, adjusts concentration of cell suspension 3 * 10
4individual/ml.
2) cell kind is entered in 96 well culture plates, every hole adds cell suspension 180 μ l, culture plate put into cell culture incubator (37 ℃, 5%C0
2) cellar culture.
3) according to Growth of Cells situation, generally grow to 50%-70%, add fukean tablet solution, continue to cultivate 24h.
4) after 24h, add 20 μ l MTT solution (5mg/ml, i.e. 0.5%MTT), continue to cultivate 4h.
5) after 4h, buckle method is removed supernatant, with absorbent paper, pats dry gently, and low-speed oscillation 10min on shaking table, as entered 200 μ l dimethyl sulfoxide, is put in every hole, and crystal is fully dissolved.At enzyme-linked immunosorbent assay instrument 490nm place, measure the light absorption value in each hole.
6) background (do not add cell, only add culture fluid) is set simultaneously, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide), sets 6 multiple holes for every group.
7) result represents the suppression ratio of cell with medicine: cell increment suppression ratio (%)=(control wells OD value-dosing holes OD value), control wells OD value * 100%.Experiment repeats 3 times.
3. statistical disposition
Adopt correlation analysis and Student t check in Microsoft Excel2007 software, data represent with mean ± S.D..
4. experimental result
Statistical result showed after mtt assay experiment, with matched group comparison, when dosage reaches 5mg/ml, to YAC-1 cell inhibitory effect variant (P<0.05), dosage this difference when 10mg/ml has significance (P<0.01), has utmost point significant difference (P<0.001) when dosage reaches 15-20mg/ml.
Table 1 fukean tablet is on YAC-1 cell inhibitory effect impact research (X ± SD)
| Group | Drug level (mg/ml) | Suppression ratio (%) |
| Matched group | 0 | 0 |
| 1 | 5 | 8.53±7.66 |
| 2 | 10 | 13.43±11.52* |
| 3 | 15 | 29.62±14.87** |
| 4 | 20 | 38.79±17.54** |
Note: with matched group comparison, * P<O.01; * P<0.001.
5. experiment conclusion
Fukean tablet can suppress YAC-1 cell proliferation, reduces the Growth of Cells number of YAC-1 cell, and this effect is dose dependent.
Claims (4)
1. the preparation method of a fukean tablet, by Herba Oxalidis strictae 1000g, Herb Polygoni Chinensis 1000g, Cortex Ilicis Rotundae 670g, Herba Plantaginis 330g, Pericarpium Granati 330g makes as crude drug, it is characterized in that, described preparation method is comprised of the following step: get upper gomi herbs, pulverize, 65% ethanol that adds 10L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600-800W, extract 2 times, each 5-10 minute, combining extraction liquid, concentrated, be added on Diaion HP-10 macroporous adsorptive resins, 65% ethanol elution, collect 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, microwave extraction thing is mixed and adds starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.3g.
2. according to the preparation method of the fukean tablet described in claim l, it is characterized in that, described microwave extracting power is 700W.
3. according to the preparation method of the fukean tablet described in claim l, it is characterized in that, the each extraction time of described microwave extracting is 8 minutes.
4. the fukean tablet described in claim l suppresses the application in mouse lymphoma cell YAC-1 cell proliferation medicine in preparation.
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| CN201310659043.6A CN103655744B (en) | 2013-12-06 | 2013-12-06 | Preparation method for Fukean tablet and application of Fukean tablet in preparation of drug used for inhibiting proliferation of mouse lymphoma cell YAC-1 |
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| CN201310659043.6A CN103655744B (en) | 2013-12-06 | 2013-12-06 | Preparation method for Fukean tablet and application of Fukean tablet in preparation of drug used for inhibiting proliferation of mouse lymphoma cell YAC-1 |
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| CN103655744B CN103655744B (en) | 2015-06-10 |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104383282A (en) * | 2014-11-13 | 2015-03-04 | 山东中大药业有限公司 | Preparation method of nine-ingredient false Chinese swertia herb tablets and application of nine-ingredient false Chinese swertia herb tablets to preparation of medicines for inhibiting proliferation of hepatoma cell Hepa1-6 |
| CN104398818A (en) * | 2014-11-13 | 2015-03-11 | 山东中大药业有限公司 | Preparation method of nine flavor swertia bimaculata tablet and application of nine flavor swertia bimaculata tablet in preparation of drugs for inhibition of cell proliferation of myeloma cell SP2/0 |
| CN104398815A (en) * | 2014-11-13 | 2015-03-11 | 山东中大药业有限公司 | Preparation method of nine flavor swertia bimaculata tablet and application of nine flavor swertia bimaculata tablet in preparation of drugs for inhibition of cell proliferation of melanoma cell B16 |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20020036807A (en) * | 2002-03-26 | 2002-05-16 | 배지현 | Plantago asiatica extraction with a function of anticancer |
| CN1887315A (en) * | 2006-05-24 | 2007-01-03 | 张嵩 | Fukean dispersible tablet and preparation method thereof |
| KR20070074763A (en) * | 2006-01-10 | 2007-07-18 | 고려대학교 산학협력단 | Extraction and purification method for effective component of plant |
| WO2011036316A2 (en) * | 2009-09-23 | 2011-03-31 | Probelte Pharma, Sa | Use of plant extracts as prebiotics, compostions and foods containing such extracts |
| CN102127142A (en) * | 2010-12-28 | 2011-07-20 | 赵全成 | Ilicis routundae cortex derivants and application thereof in preparing medicament capable of resisting tumors |
| KR101099718B1 (en) * | 2008-12-26 | 2011-12-28 | 고흥석류친환경영농조합법인 | Composition for prevention and treatment of obesity comprising Punica granatum Extracts |
| CN102824409A (en) * | 2012-09-24 | 2012-12-19 | 南京正亮医药科技有限公司 | Preparation method of liver protection tablets |
| CN102836256A (en) * | 2012-09-24 | 2012-12-26 | 南京正亮医药科技有限公司 | Preparation method for baipuhuang tablet (traditional Chinese medicine tablet) |
-
2013
- 2013-12-06 CN CN201310659043.6A patent/CN103655744B/en not_active Expired - Fee Related
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20020036807A (en) * | 2002-03-26 | 2002-05-16 | 배지현 | Plantago asiatica extraction with a function of anticancer |
| KR20070074763A (en) * | 2006-01-10 | 2007-07-18 | 고려대학교 산학협력단 | Extraction and purification method for effective component of plant |
| CN1887315A (en) * | 2006-05-24 | 2007-01-03 | 张嵩 | Fukean dispersible tablet and preparation method thereof |
| KR101099718B1 (en) * | 2008-12-26 | 2011-12-28 | 고흥석류친환경영농조합법인 | Composition for prevention and treatment of obesity comprising Punica granatum Extracts |
| WO2011036316A2 (en) * | 2009-09-23 | 2011-03-31 | Probelte Pharma, Sa | Use of plant extracts as prebiotics, compostions and foods containing such extracts |
| CN102127142A (en) * | 2010-12-28 | 2011-07-20 | 赵全成 | Ilicis routundae cortex derivants and application thereof in preparing medicament capable of resisting tumors |
| CN102824409A (en) * | 2012-09-24 | 2012-12-19 | 南京正亮医药科技有限公司 | Preparation method of liver protection tablets |
| CN102836256A (en) * | 2012-09-24 | 2012-12-26 | 南京正亮医药科技有限公司 | Preparation method for baipuhuang tablet (traditional Chinese medicine tablet) |
Non-Patent Citations (3)
| Title |
|---|
| 徐国钧等: "《抗肿瘤中草药彩色图谱》", 30 June 1997, article "石榴皮" * |
| 蔡明坤: "《临症中医视角：内科常见病辑》", 31 January 2012, article "肿瘤", pages: 354-355 * |
| 黄红兵: "《抗肿瘤中药临床应用与图谱》", 31 January 2008, article "车前草", pages: 62-63 - 273 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104383282A (en) * | 2014-11-13 | 2015-03-04 | 山东中大药业有限公司 | Preparation method of nine-ingredient false Chinese swertia herb tablets and application of nine-ingredient false Chinese swertia herb tablets to preparation of medicines for inhibiting proliferation of hepatoma cell Hepa1-6 |
| CN104398818A (en) * | 2014-11-13 | 2015-03-11 | 山东中大药业有限公司 | Preparation method of nine flavor swertia bimaculata tablet and application of nine flavor swertia bimaculata tablet in preparation of drugs for inhibition of cell proliferation of myeloma cell SP2/0 |
| CN104398815A (en) * | 2014-11-13 | 2015-03-11 | 山东中大药业有限公司 | Preparation method of nine flavor swertia bimaculata tablet and application of nine flavor swertia bimaculata tablet in preparation of drugs for inhibition of cell proliferation of melanoma cell B16 |
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